基于核酸扩增技术-3D石墨烯高灵敏、免标记和普适性电化学检测海水与海产品中食源性弧菌

项目名称： 基于核酸扩增技术-3D石墨烯高灵敏、免标记和普适性电化学检测海水与海产品中食源性弧菌

项目编号： No.41476083

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 天文学、地球科学

项目作者： 杨涛

作者单位： 中山大学

项目金额： 86万元

中文摘要： 本项目拟将3D石墨烯、核酸扩增技术和电化学DNA传感器技术相结合，开展海水及海产品中食源性病原弧菌高灵敏、免标记和普适性电化学检测研究。设计一系列3D石墨烯及其复合材料传感界面，对比各种制备方案对界面形貌、电化学信号等的影响。借助传感界面受DNA固定与杂交的影响而产生界面电化学信号的变化（信号关或信号开），成功实现对DNA探针的固定及目标扩增产物的杂交识别。传感界面优异的导电性、丰富的边缘缺陷，高的比表面积与强的机械性能等优点增强纳米界面的电化学响应与稳定性，结合核酸扩增技术，提高检测灵敏度。各种电化学活性功能团，丰富的共轭结构和DNA碱基之间的非共价π-π堆积作用，确保无损、免标记与普适性检测。比较不同传感界面的组成、尺度等因素对电极稳定性，探针固定效率、灵敏度以及再生性能的影响。有效整合核酸扩增模块与电化学检测模块，最终实现高灵敏、免标记、普适性检测多种食源性弧菌。

中文关键词： 海洋监测；核酸扩增技术；3D石墨烯；食源性弧菌；电化学生物传感器

英文摘要： In this project, novel 3D graphene will be integrated with nucleic acid amplification technique and electrochemical biosensor for highly sensitive, label-free and universal detection of vibrios originated from seawater and seafoods. Physical and chemical routes will be adopted to prepare a series of 3D graphene sensing platforms. We will study the effect of different preparation schemes and experimental parameters on the morphology, size, and electrochemical signals of the sensing platforms. The sensing platforms possess plenty of electrochemical sites, high specific surface area, powerful mechanical properties, etc, which endow sensing platforms with super conductivity, highly electrochemical activities and stability in different interfaces and phases. The immobilization of the probe will be mainly conducted via covalent or noncovalent methods for universal detection. The noncovalent method depends on the π-π interaction between rich-conjugated structures on 3D graphene sensing platform and DNA bases. The reversible specific covalent link between phenylboronic acid and 1,2-hydroxy functinal groups also insures the freely switchable tests. In the presence of target, the binding hybrids will disturb the interaction between probes and 3D graphene, accompanied with the electrochemical signal change of the nanointerfaces (signal-on and signal-off). The signal change will be served as a powerful tool to investigate the effect of different morphologies and sizes of sensing platforms on the loading of the probe, sensitivity, recovery of sensing platform. The synergistic effect for improving the sensitivity of the target detection will be pursued. Above 3D graphene sensing platforms will be integrated with nucleic acid amplification technique to highly sensitively and universally detect the gene sequences originated from real sample of the vibrios originated from seawater and seafoods.

英文关键词： marine monotoring;nucleic acid amplification technique;3D graphene;foodborn vibrio;electrochemical biosensor